Autoinducer of virulence as a target for vaccine and therapy against Staphylococcus aureus

Science

Volumn 280, Issue 5362, 1998, Pages 438-440

  Balaban, Naomi ^a   Goldkorn, Tzipora ^b   Nhan, Rachael T ^a   Dang, Luong B ^b   Scott, Steven ^a   Ridgley, Rose M ^a   Rasooly, Avraham ^c   Wright, Susan C ^d   Larrick, James W ^d   Rasooly, Reuven ^a   Carlson, James R ^a  

^a UNIVERSITY OF CALIFORNIA   (United States)

^b UNIVERSITY OF CALIFORNIA   (United States)

^c UNIVERSITY OF MARYLAND   (United States)

^d Palo Alto Institute of Molecular Medicine   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

BACTERIAL PROTEIN; RNA III ACTIVATING PROTEIN; RNA III INHIBITING PEPTIDE; STAPHYLOCOCCUS VACCINE; UNCLASSIFIED DRUG;

ANIMAL EXPERIMENT; ANIMAL MODEL; ARTICLE; CONTROLLED STUDY; HOST RESISTANCE; INFECTION PREVENTION; MOUSE; NONHUMAN; PRIORITY JOURNAL; STAPHYLOCOCCUS AUREUS; STAPHYLOCOCCUS INFECTION; VACCINATION;

AMINO ACID SEQUENCE; ANIMALS; ANTIBODIES, BACTERIAL; BACTERIAL PROTEINS; BACTERIAL TOXINS; BACTERIAL VACCINES; MALE; MICE; MICE, INBRED HRS; OLIGOPEPTIDES; RNA, ANTISENSE; RNA, BACTERIAL; SIGNAL TRANSDUCTION; STAPHYLOCOCCAL SKIN INFECTIONS; STAPHYLOCOCCUS AUREUS; VACCINATION; VIRULENCE;

ANIMALIA; BACTERIA (MICROORGANISMS); POSIBACTERIA; STAPHYLOCOCCUS AUREUS;

EID: 0032540422     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.280.5362.438     Document Type: Article

References (28)

1. S. C. Silverstein and T. H. Steinberg, in Microbiology, B. D. Davis, R. Dulbecco, H. N. Eisen, H. S. Ginsberg, Eds. (Lippincott, Philadelphia, 1990), pp. 485-506.
2. A. L. Panlilio et al., Infect Control Hosp. Epidemiol. 13, 582 (1992).
3. R. P. Novick et al., EMBO J. 12, 3967 (1993).
4. N. Balaban and R. P. Novick, FEMS Microbiol. Lett. 133, 155 (1995).
5. E. Moerfeldt, D. Taylor, A. Von Gabain, S. Arvidson, EMBO J. 14, 4569 (1995).
6. N. Balaban and R. P. Novick, Proc. Natl. Acad. Sci. U.S.A. 92, 1619 (1995).
7. Bacterial strains used were wild-type S. aureus; nonpathogenic agr-null S. aureus in which the whole 3.4-kb agr fragment was replaced by the tetM marker (3); nonpathogenic strain of S. aureus producing RIP (6); and SD, a highly encapsulated strain of S. aureus (8) that produces exopolysaccharides that are antigenically identical to many clinical S. aureus strains tested (27).
8. C. Bunce, L. Wheeler, G. Reed, J. Musser, N. Barg, Infect. Immun. 60, 2636 (1992).
9. Animals [4-to 8-week-old (20 to 30 g) outbred, immunocompetent hairless male mice, strain Crl: SKH1(hrhr)Br] were obtained from Charles River Laboratories (Wilmington, MA).
10. 8 SD S. aureus subcutaneously together with 1 mg of Cytodex beads (10) to induce a local infection. The size of the lesion was measured daily (14). Of note is that animals injected only with Cytodex beads developed no lesions at all.
11. Fisher's exact probability test was used to compare proportions of mice developing lesions and mice developing RAP antibodies among the experimental groups (RAP vaccinated, CFA controls, untreated controls). Among animals that developed lesions after challenge with S. aureus, the size of the lesions was compared by single-factor analysis of variance. Post-hoc testing was done by Fisher's protected least significant difference.
12. Lesions were measured 24 to 48 hours after challenge and are presented as area = 0.5[π(length) × (width)].
13. To estimate amount of antibody to the injected antigen, we collected a drop of blood (50 μl) from the tip of the tail before vaccination (preimmune sera) and 7 days after the third vaccination period (postimmune sera, 3 days before bacterial challenge). RAP antibody titer was determined by Western blotting. Serum was added to the blot (containing postexponential supernatants) in increasing dilutions until no band appeared. The highest dilution that still reacted with RAP is the titer determined.
14. For Western blotting, 30 μl of wild-type S. aureus (concentrated 10:1) postexponential supernatant or 5 μg of purified RAP was separated on a 12% SDS-polyacrylamide gel and blotted onto a nitrocellulose membrane; the membrane was blocked for 1 hour in 3% bovine serum albumin (BSA) in tris-buffered saline (TBS). The membrane was then incubated with the serum (diluted in 1% BSA/TBS) for 2 hours at 22°C, washed three times for 10 min in TBS containing 0.05% Tween 20 (TBST), and incubated in peroxidase-conjugated mouse immunoglobulin G (Boehringer) for 1 hour at 37°C in TBST. Bound antibody was detected by chemiluminescence with the ECL Western blotting system (Amersham).
15. Single-letter abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; U, Tyr; X, unknown or other.
16. 9) were incubated in the presence of RIP that was purified from 5 ml of postexponential culture broth in saline (6), or with saline only as a control, with 0.5 mg of synthetic RIP (Pep) in a final dimethyl sulfoxide (DMSO) solution of 3% (in saline), or with 3% DMSO only in saline as a control, for 30 min at 37°C. The bacteria-RIP, bacteria-Pep, bacteria-saline, or bacteria-DMSO mixture was injected subcutaneously with Cytodex beads (1 mg) (8) into 8-week-old immunocompetent hairless male mice to induce a local infection. The lesion was measured daily (12).
17. J. C. Lee, Trends Microbiol. 4, 162 (1996).
18. R. Naso and A. Fattom, in Novel Strategies in Design and Production of Vaccines, S. Cohen and A. Shafferman, Eds. (Plenum, New York, 1996), pp. 133-140.
19. M. Arthur and P. Courvalin, Antimicrob. Agents Chemother. 37, 1563 (1993).
20. W. C. Noble, Z. Virani, R.G. Cree, FEMS Microbiol. Lett. 93, 195 (1992).
21. J. Guangyong, R. Beavis, R. P. Novick, Proc. Natl. Acad. Sci. U.S.A. 92, 12055 (1995).
22. R. Rappuoli, V. Scarlato, B. Arico, N. Balaban, in Signal Transduction and Bacterial Virulence, R. Rappuoli, V. Scarlato, B. Arico, Eds. (Springer Verlag, Heidelberg, 1995), pp. 1-4.
23. R. Magnuson, J. Solomon, A. D. Grossman, Cell 77, 207 (1994).
24. S. Swift, J. N. Bainton, M. K. Winson, Trends Microbiol. 2, 193 (1994).
25. J. P. Pearson, L. Passador, B. Iglewski, E. P. Greenberg, Proc. Natl. Acad. Sci. U.S.A. 92, 1490 (1995).
26. S. Roychoudhury et al., ibid. 90, 965 (1993).
27. T. E. West, M. E. West, J. M. Mylotte, J. Clin. Microbiol. 21, 490 (1985).
28. We thank N. Lerche and P. Kass for statistical analysis, A. Moses for helpful discussions, B. Menzies for kindly providing us with the SD S. aureus strain, M. Balaban for reviewing the article, and D. Schwartz for his endless support. Supported by grants from NIH (AI40830), CTR, the American Heart Association, and UC Davis Dean's Research Fund.